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AQ inhibits Plasmodium falciparum phosphoethanolamine methyltransferase, an enzyme found in malarial parasites but not in humans 43. Here we report that AQ also binds and blocks the function of host cathepsin B protein and thus entry of key pathogens into the cytoplasm. Since AQ has been used for the past 60 years, its safety and pharmacokinetic profiles are well known. For example, it has been determined that after oral administration in healthy subjects, AQ is quickly absorbed and biotransformed into its main active form, DEAQ, which in turn is slowly eliminated with a terminal half-life of 9’18 days. The availability of PK values for AQ allows us to compare it to our in vitro data. The EC50 values for all of the tested pathogens inhibited by AQ or by DEAQ were at low ?M range. Importantly, the known Cmax values (maximal amounts of drug in the blood after oral administration) of DEAQ in humans are in low ?M range 43,44,45 and are comparable to the DEAQ EC50 values found in vitro for tested pathogens. This suggests that it is possible to achieve anti-pathogen effective AQ doses in blood by orally administered AQ. All toxins were purchased from List Biological Laboratories. FP59 was a kind gift from S. Leppla (National Institute of Allergy and Infectious Diseases/National Institutes of Health, Bethesda, MD). An FDA-approved drug library comprising of 1,581 drugs was purchased from Johns Hopkins, titled, Johns Hopkins Clinical Compound Library (JHCCL) version 1. The drugs arrived as 10 mM stock solutions in sealed microtiter plates and were made using DMSO or water as solvents. Drugs were arrayed in 96-well plates and screened at a stock concentration of 10 mM. The library was stored at ?20 ‘C until use. Prior to use, the library of drugs was thawed at 25 ‘C. Compounds that were determined to be compounds of interest, were isolated and reproduced from 10 mM solutions. Amodiaquine, Chloroquine, Cinchonine, Primaquine and Quinidine were purchased from Sigma-Aldrich (St. All drugs were prepared at 10 mM using DMSO as a solvent. Desethyl-amodiaquine and desethyl-chloroquine were purchased from Toronto Research Chemicals Inc. Anti’N-terminal MEK-2, anti-tubulin and anti-PA antibodies were purchased from Santa Cruz Biotechnology. Purified human cathepsin B protein used for NMR experiments was purchased from ACROBiosystems, legal steroids reviews. During the drug library screen assay, 0. Cells were treated with compounds for 1 hour at 37 ‘C 5%CO 2 and then challenged with anthrax toxins, such that the final toxins concentrations were 0. Cells were treated with toxin and drugs for 6 hours.
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At large doses of exogenous androgens, spermatogenesis may also be suppressed through feedback inhibition of pituitary follicle stimulating hormone (FSH). There is a lack of substantial evidence that androgens are effective in fractures, surgery, convalescence, and functional uterine bleeding, legal steroids price
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